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Simultaneous CRISPR/Cas9 Editing of Three PPO Genes Reduces Fruit Flesh Browning in Solanum melongena L.

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Simultaneous CRISPR/Cas9 Editing of Three PPO Genes Reduces Fruit Flesh Browning in Solanum melongena L.

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dc.contributor.author Maioli, Alex es_ES
dc.contributor.author Gianoglio, Silvia es_ES
dc.contributor.author Moglia, Andrea es_ES
dc.contributor.author Acquadro, Alberto es_ES
dc.contributor.author Valentino, Danila es_ES
dc.contributor.author Milani, Anna Maria es_ES
dc.contributor.author Prohens Tomás, Jaime es_ES
dc.contributor.author Orzáez Calatayud, Diego Vicente es_ES
dc.contributor.author GRANELL RICHART, ANTONIO es_ES
dc.contributor.author Lanteri, Sergio es_ES
dc.contributor.author Comino, Cinzia es_ES
dc.date.accessioned 2021-06-15T03:31:35Z
dc.date.available 2021-06-15T03:31:35Z
dc.date.issued 2020-12-03 es_ES
dc.identifier.uri http://hdl.handle.net/10251/167985
dc.description.abstract [EN] Polyphenol oxidases (PPOs) catalyze the oxidization of polyphenols, which in turn causes the browning of the eggplant berry flesh after cutting. This has a negative impact on fruit quality for both industrial transformation and fresh consumption. Ten PPO genes (named SmelPPO1-10) were identified in eggplant thanks to the recent availability of a high-quality genome sequence. A CRISPR/Cas9-based mutagenesis approach was applied to knock-out three target PPO genes (SmelPPO4, SmelPPO5, and SmelPPO6), which showed high transcript levels in the fruit after cutting. An optimized transformation protocol for eggplant cotyledons was used to obtain plants in which Cas9 is directed to a conserved region shared by the three PPO genes. The successful editing of the SmelPPO4, SmelPPO5, and SmelPPO6 loci of in vitro regenerated plantlets was confirmed by Illumina deep sequencing of amplicons of the target sites. Besides, deep sequencing of amplicons of the potential off-target loci identified in silico proved the absence of detectable non-specific mutations. The induced mutations were stably inherited in the T-1 and T-2 progeny and were associated with a reduced PPO activity and browning of the berry flesh after cutting. Our results provide the first example of the use of the CRISPR/Cas9 system in eggplant for biotechnological applications and open the way to the development of eggplant genotypes with low flesh browning which maintain a high polyphenol content in the berries. es_ES
dc.description.sponsorship Research was financially supported by the project CRISPR/Cas9-mediated gene knock-out in eggplant financed by Compagnia San Paolo. es_ES
dc.language Inglés es_ES
dc.publisher Frontiers Media SA es_ES
dc.relation.ispartof Frontiers in Plant Science es_ES
dc.rights Reconocimiento (by) es_ES
dc.subject Gene editing es_ES
dc.subject CRISPR es_ES
dc.subject Cas 9 es_ES
dc.subject Eggplant es_ES
dc.subject Polyphenol oxydase es_ES
dc.subject Knock-out es_ES
dc.subject.classification GENETICA es_ES
dc.subject.classification BIOQUIMICA Y BIOLOGIA MOLECULAR es_ES
dc.title Simultaneous CRISPR/Cas9 Editing of Three PPO Genes Reduces Fruit Flesh Browning in Solanum melongena L. es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.3389/fpls.2020.607161 es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia es_ES
dc.description.bibliographicCitation Maioli, A.; Gianoglio, S.; Moglia, A.; Acquadro, A.; Valentino, D.; Milani, AM.; Prohens Tomás, J.... (2020). Simultaneous CRISPR/Cas9 Editing of Three PPO Genes Reduces Fruit Flesh Browning in Solanum melongena L. Frontiers in Plant Science. 11:1-13. https://doi.org/10.3389/fpls.2020.607161 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion https://doi.org/10.3389/fpls.2020.607161 es_ES
dc.description.upvformatpinicio 1 es_ES
dc.description.upvformatpfin 13 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 11 es_ES
dc.identifier.eissn 1664-462X es_ES
dc.identifier.pmid 33343607 es_ES
dc.identifier.pmcid PMC7744776 es_ES
dc.relation.pasarela S\431064 es_ES
dc.contributor.funder Compagnia di San Paolo es_ES
dc.description.references Andersson, M., Turesson, H., Nicolia, A., Fält, A.-S., Samuelsson, M., & Hofvander, P. (2016). Efficient targeted multiallelic mutagenesis in tetraploid potato (Solanum tuberosum) by transient CRISPR-Cas9 expression in protoplasts. Plant Cell Reports, 36(1), 117-128. doi:10.1007/s00299-016-2062-3 es_ES
dc.description.references Bachem, C. W. B., Speckmann, G.-J., van der Linde, P. C. G., Verheggen, F. T. M., Hunt, M. D., Steffens, J. C., & Zabeau, M. (1994). Antisense Expression of Polyphenol Oxidase Genes Inhibits Enzymatic Browning in Potato Tubers. Bio/Technology, 12(11), 1101-1105. doi:10.1038/nbt1194-1101 es_ES
dc.description.references Barchi, L., Pietrella, M., Venturini, L., Minio, A., Toppino, L., Acquadro, A., … Rotino, G. L. (2019). A chromosome-anchored eggplant genome sequence reveals key events in Solanaceae evolution. Scientific Reports, 9(1). doi:10.1038/s41598-019-47985-w es_ES
dc.description.references Bellés, J. M., Garro, R., Pallás, V., Fayos, J., Rodrigo, I., & Conejero, V. (2005). Accumulation of gentisic acid as associated with systemic infections but not with the hypersensitive response in plant-pathogen interactions. Planta, 223(3), 500-511. doi:10.1007/s00425-005-0109-8 es_ES
dc.description.references Bolger, A. M., Lohse, M., & Usadel, B. (2014). Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics, 30(15), 2114-2120. doi:10.1093/bioinformatics/btu170 es_ES
dc.description.references Bortesi, L., Zhu, C., Zischewski, J., Perez, L., Bassié, L., Nadi, R., … Schillberg, S. (2016). Patterns of CRISPR/Cas9 activity in plants, animals and microbes. Plant Biotechnology Journal, 14(12), 2203-2216. doi:10.1111/pbi.12634 es_ES
dc.description.references Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2), 248-254. doi:10.1016/0003-2697(76)90527-3 es_ES
dc.description.references Chi, M., Bhagwat, B., Lane, W. D., Tang, G., Su, Y., Sun, R., … Xiang, Y. (2014). Reduced polyphenol oxidase gene expression and enzymatic browning in potato (Solanum tuberosum L.) with artificial microRNAs. BMC Plant Biology, 14(1). doi:10.1186/1471-2229-14-62 es_ES
dc.description.references Clement, K., Rees, H., Canver, M. C., Gehrke, J. M., Farouni, R., Hsu, J. Y., … Pinello, L. (2019). CRISPResso2 provides accurate and rapid genome editing sequence analysis. Nature Biotechnology, 37(3), 224-226. doi:10.1038/s41587-019-0032-3 es_ES
dc.description.references Coetzer, C., Corsini, D., Love, S., Pavek, J., & Tumer, N. (2001). Control of Enzymatic Browning in Potato (Solanum tuberosum L.) by Sense and Antisense RNA from Tomato Polyphenol Oxidase. Journal of Agricultural and Food Chemistry, 49(2), 652-657. doi:10.1021/jf001217f es_ES
dc.description.references Cong, L., & Zhang, F. (2014). Genome Engineering Using CRISPR-Cas9 System. Methods in Molecular Biology, 197-217. doi:10.1007/978-1-4939-1862-1_10 es_ES
dc.description.references Docimo, T., Francese, G., De Palma, M., Mennella, D., Toppino, L., Lo Scalzo, R., … Tucci, M. (2016). Insights in the Fruit Flesh Browning Mechanisms in Solanum melongena Genetic Lines with Opposite Postcut Behavior. Journal of Agricultural and Food Chemistry, 64(22), 4675-4685. doi:10.1021/acs.jafc.6b00662 es_ES
dc.description.references Doudna, J. A., & Charpentier, E. (2014). The new frontier of genome engineering with CRISPR-Cas9. Science, 346(6213). doi:10.1126/science.1258096 es_ES
dc.description.references Feng, Z., Zhang, B., Ding, W., Liu, X., Yang, D.-L., Wei, P., … Zhu, J.-K. (2013). Efficient genome editing in plants using a CRISPR/Cas system. Cell Research, 23(10), 1229-1232. doi:10.1038/cr.2013.114 es_ES
dc.description.references Fu, Y., Foden, J. A., Khayter, C., Maeder, M. L., Reyon, D., Joung, J. K., & Sander, J. D. (2013). High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology, 31(9), 822-826. doi:10.1038/nbt.2623 es_ES
dc.description.references Gao, J., Wang, G., Ma, S., Xie, X., Wu, X., Zhang, X., … Xia, Q. (2014). CRISPR/Cas9-mediated targeted mutagenesis in Nicotiana tabacum. Plant Molecular Biology, 87(1-2), 99-110. doi:10.1007/s11103-014-0263-0 es_ES
dc.description.references García-Fortea, E., Lluch-Ruiz, A., Pineda-Chaza, B. J., García-Pérez, A., Bracho-Gil, J. P., Plazas, M., … Prohens, J. (2020). A highly efficient organogenesis protocol based on zeatin riboside for in vitro regeneration of eggplant. BMC Plant Biology, 20(1). doi:10.1186/s12870-019-2215-y es_ES
dc.description.references González, M. N., Massa, G. A., Andersson, M., Turesson, H., Olsson, N., Fält, A.-S., … Feingold, S. E. (2020). Reduced Enzymatic Browning in Potato Tubers by Specific Editing of a Polyphenol Oxidase Gene via Ribonucleoprotein Complexes Delivery of the CRISPR/Cas9 System. Frontiers in Plant Science, 10. doi:10.3389/fpls.2019.01649 es_ES
dc.description.references Guri, A., & Sink, K. C. (1988). Agrobacterium Transformation of Eggplant. Journal of Plant Physiology, 133(1), 52-55. doi:10.1016/s0176-1617(88)80083-x es_ES
dc.description.references Hahn, F., & Nekrasov, V. (2018). CRISPR/Cas precision: do we need to worry about off-targeting in plants? Plant Cell Reports, 38(4), 437-441. doi:10.1007/s00299-018-2355-9 es_ES
dc.description.references Jukanti, A. K., & Bhatt, R. (2014). Eggplant (Solanum melongena L.) polyphenol oxidase multi-gene family: a phylogenetic evaluation. 3 Biotech, 5(1), 93-99. doi:10.1007/s13205-014-0195-z es_ES
dc.description.references Karunarathna, N. L., Wang, H., Harloff, H., Jiang, L., & Jung, C. (2020). Elevating seed oil content in a polyploid crop by induced mutations in SEED FATTY ACID REDUCER genes. Plant Biotechnology Journal, 18(11), 2251-2266. doi:10.1111/pbi.13381 es_ES
dc.description.references Kaushik, P., Gramazio, P., Vilanova, S., Raigón, M. D., Prohens, J., & Plazas, M. (2017). Phenolics content, fruit flesh colour and browning in cultivated eggplant, wild relatives and interspecific hybrids and implications for fruit quality breeding. Food Research International, 102, 392-401. doi:10.1016/j.foodres.2017.09.028 es_ES
dc.description.references Li, L., & Steffens, J. (2002). Overexpression of polyphenol oxidase in transgenic tomato plants results in enhanced bacterial disease resistance. Planta, 215(2), 239-247. doi:10.1007/s00425-002-0750-4 es_ES
dc.description.references Li, Z., Liu, Z.-B., Xing, A., Moon, B. P., Koellhoffer, J. P., Huang, L., … Cigan, A. M. (2015). Cas9-Guide RNA Directed Genome Editing in Soybean. Plant Physiology, 169(2), 960-970. doi:10.1104/pp.15.00783 es_ES
dc.description.references Llorente, B., Alonso, G. D., Bravo-Almonacid, F., Rodríguez, V., López, M. G., Carrari, F., … Flawiá, M. M. (2011). Safety assessment of nonbrowning potatoes: opening the discussion about the relevance of substantial equivalence on next generation biotech crops. Plant Biotechnology Journal, 9(2), 136-150. doi:10.1111/j.1467-7652.2010.00534.x es_ES
dc.description.references Llorente, B., López, M. G., Carrari, F., Asís, R., Di Paola Naranjo, R. D., Flawiá, M. M., … Bravo-Almonacid, F. (2014). Downregulation of polyphenol oxidase in potato tubers redirects phenylpropanoid metabolism enhancing chlorogenate content and late blight resistance. Molecular Breeding, 34(4), 2049-2063. doi:10.1007/s11032-014-0162-8 es_ES
dc.description.references Ma, X., Zhang, Q., Zhu, Q., Liu, W., Chen, Y., Qiu, R., … Liu, Y.-G. (2015). A Robust CRISPR/Cas9 System for Convenient, High-Efficiency Multiplex Genome Editing in Monocot and Dicot Plants. Molecular Plant, 8(8), 1274-1284. doi:10.1016/j.molp.2015.04.007 es_ES
dc.description.references Mahanil, S., Attajarusit, J., Stout, M. J., & Thipyapong, P. (2008). Overexpression of tomato polyphenol oxidase increases resistance to common cutworm. Plant Science, 174(4), 456-466. doi:10.1016/j.plantsci.2008.01.006 es_ES
dc.description.references Menin, B., Moglia, A., Comino, C., Hakkert, J. C., Lanteri, S., & Beekwilder, J. (2013). In vitrocallus-induction in globe artichoke (Cynara cardunculusL. var.scolymus) as a system for the production of caffeoylquinic acids. The Journal of Horticultural Science and Biotechnology, 88(5), 537-542. doi:10.1080/14620316.2013.11513003 es_ES
dc.description.references Mennella, G., Lo Scalzo, R., Fibiani, M., D’Alessandro, A., Francese, G., Toppino, L., … Rotino, G. L. (2012). Chemical and Bioactive Quality Traits During Fruit Ripening in Eggplant (S. melongena L.) and Allied Species. Journal of Agricultural and Food Chemistry, 60(47), 11821-11831. doi:10.1021/jf3037424 es_ES
dc.description.references Miao, J., Guo, D., Zhang, J., Huang, Q., Qin, G., Zhang, X., … Qu, L.-J. (2013). Targeted mutagenesis in rice using CRISPR-Cas system. Cell Research, 23(10), 1233-1236. doi:10.1038/cr.2013.123 es_ES
dc.description.references Mishra, B. B., Gautam, S., & Sharma, A. (2013). Free phenolics and polyphenol oxidase (PPO): The factors affecting post-cut browning in eggplant (Solanum melongena). Food Chemistry, 139(1-4), 105-114. doi:10.1016/j.foodchem.2013.01.074 es_ES
dc.description.references Muktadir, M. A., Habib, M. A., Khaleque Mian, M. A., & Yousuf Akhond, M. A. (2016). Regeneration efficiency based on genotype, culture condition and growth regulators of eggplant (Solanum melongena L.). Agriculture and Natural Resources, 50(1), 38-42. doi:10.1016/j.anres.2014.10.001 es_ES
dc.description.references Naveed, M., Hejazi, V., Abbas, M., Kamboh, A. A., Khan, G. J., Shumzaid, M., … XiaoHui, Z. (2018). Chlorogenic acid (CGA): A pharmacological review and call for further research. Biomedicine & Pharmacotherapy, 97, 67-74. doi:10.1016/j.biopha.2017.10.064 es_ES
dc.description.references Nonaka, S., Arai, C., Takayama, M., Matsukura, C., & Ezura, H. (2017). Efficient increase of ɣ-aminobutyric acid (GABA) content in tomato fruits by targeted mutagenesis. Scientific Reports, 7(1). doi:10.1038/s41598-017-06400-y es_ES
dc.description.references Pan, C., Ye, L., Qin, L., Liu, X., He, Y., Wang, J., … Lu, G. (2016). CRISPR/Cas9-mediated efficient and heritable targeted mutagenesis in tomato plants in the first and later generations. Scientific Reports, 6(1). doi:10.1038/srep24765 es_ES
dc.description.references Peterson, B. A., Haak, D. C., Nishimura, M. T., Teixeira, P. J. P. L., James, S. R., Dangl, J. L., & Nimchuk, Z. L. (2016). Genome-Wide Assessment of Efficiency and Specificity in CRISPR/Cas9 Mediated Multiple Site Targeting in Arabidopsis. PLOS ONE, 11(9), e0162169. doi:10.1371/journal.pone.0162169 es_ES
dc.description.references Plazas, M., López-Gresa, M. P., Vilanova, S., Torres, C., Hurtado, M., Gramazio, P., … Prohens, J. (2013). Diversity and Relationships in Key Traits for Functional and Apparent Quality in a Collection of Eggplant: Fruit Phenolics Content, Antioxidant Activity, Polyphenol Oxidase Activity, and Browning. Journal of Agricultural and Food Chemistry, 61(37), 8871-8879. doi:10.1021/jf402429k es_ES
dc.description.references Prohens, J., Rodríguez-Burruezo, A., Raigón, M. D., & Nuez, F. (2007). Total Phenolic Concentration and Browning Susceptibility in a Collection of Different Varietal Types and Hybrids of Eggplant: Implications for Breeding for Higher Nutritional Quality and Reduced Browning. Journal of the American Society for Horticultural Science, 132(5), 638-646. doi:10.21273/jashs.132.5.638 es_ES
dc.description.references Rommens, C. M., Ye, J., Richael, C., & Swords, K. (2006). Improving Potato Storage and Processing Characteristics through All-Native DNA Transformation. Journal of Agricultural and Food Chemistry, 54(26), 9882-9887. doi:10.1021/jf062477l es_ES
dc.description.references Rotino, G. L., Sala, T., & Toppino, L. (2013). Eggplant. Alien Gene Transfer in Crop Plants, Volume 2, 381-409. doi:10.1007/978-1-4614-9572-7_16 es_ES
dc.description.references Saini, D. K., & Kaushik, P. (2019). Visiting eggplant from a biotechnological perspective: A review. Scientia Horticulturae, 253, 327-340. doi:10.1016/j.scienta.2019.04.042 es_ES
dc.description.references Sashidhar, N., Harloff, H. J., Potgieter, L., & Jung, C. (2020). Gene editing of three BnITPK genes in tetraploid oilseed rape leads to significant reduction of phytic acid in seeds. Plant Biotechnology Journal, 18(11), 2241-2250. doi:10.1111/pbi.13380 es_ES
dc.description.references Shetty, S. M., Chandrashekar, A., & Venkatesh, Y. P. (2011). Eggplant polyphenol oxidase multigene family: Cloning, phylogeny, expression analyses and immunolocalization in response to wounding. Phytochemistry, 72(18), 2275-2287. doi:10.1016/j.phytochem.2011.08.028 es_ES
dc.description.references Svitashev, S., Young, J. K., Schwartz, C., Gao, H., Falco, S. C., & Cigan, A. M. (2015). Targeted Mutagenesis, Precise Gene Editing, and Site-Specific Gene Insertion in Maize Using Cas9 and Guide RNA. Plant Physiology, 169(2), 931-945. doi:10.1104/pp.15.00793 es_ES
dc.description.references Taranto, F., Pasqualone, A., Mangini, G., Tripodi, P., Miazzi, M., Pavan, S., & Montemurro, C. (2017). Polyphenol Oxidases in Crops: Biochemical, Physiological and Genetic Aspects. International Journal of Molecular Sciences, 18(2), 377. doi:10.3390/ijms18020377 es_ES
dc.description.references Thipyapong, P., Hunt, M. D., & Steffens, J. C. (2004). Antisense downregulation of polyphenol oxidase results in enhanced disease susceptibility. Planta, 220(1), 105-117. doi:10.1007/s00425-004-1330-6 es_ES
dc.description.references Thipyapong, P., Joel, D. M., & Steffens, J. C. (1997). Differential Expression and Turnover of the Tomato Polyphenol Oxidase Gene Family during Vegetative and Reproductive Development. Plant Physiology, 113(3), 707-718. doi:10.1104/pp.113.3.707 es_ES
dc.description.references Van Eck, J. (2018). Genome editing and plant transformation of solanaceous food crops. Current Opinion in Biotechnology, 49, 35-41. doi:10.1016/j.copbio.2017.07.012 es_ES
dc.description.references Wang, Z.-P., Xing, H.-L., Dong, L., Zhang, H.-Y., Han, C.-Y., Wang, X.-C., & Chen, Q.-J. (2015). Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biology, 16(1). doi:10.1186/s13059-015-0715-0 es_ES
dc.description.references Wolt, J. D., Wang, K., & Yang, B. (2015). The Regulatory Status of Genome‐edited Crops. Plant Biotechnology Journal, 14(2), 510-518. doi:10.1111/pbi.12444 es_ES
dc.description.references Zhang, Q., Xing, H.-L., Wang, Z.-P., Zhang, H.-Y., Yang, F., Wang, X.-C., & Chen, Q.-J. (2018). Potential high-frequency off-target mutagenesis induced by CRISPR/Cas9 in Arabidopsis and its prevention. Plant Molecular Biology, 96(4-5), 445-456. doi:10.1007/s11103-018-0709-x es_ES
dc.description.references Zheng, N., Li, T., Dittman, J. D., Su, J., Li, R., Gassmann, W., … Yang, B. (2020). CRISPR/Cas9-Based Gene Editing Using Egg Cell-Specific Promoters in Arabidopsis and Soybean. Frontiers in Plant Science, 11. doi:10.3389/fpls.2020.00800 es_ES


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